The International Journal of Biochemistry & Cell Biology 36 (2004) 1367–1373

Cells in focus
Odontoblasts: the cells forming and maintaining dentine
Victor E. Arana-Chavez∗ , Luciana F. Massa
Laboratory of Mineralized Tissue Biology, Department of Histology and Embryology, Institute of Biomedical Sciences,
University of São Paulo, Av. Prof. Lineu Prestes, 1524 Cidade Universitaria, 05508-900 São Paulo, SP, Brazil
Received 13 September 2003; received in revised form 19 December 2003; accepted 13 January 2004

Abstract
Odontoblasts are tall columnar cells located at the periphery of the dental pulp. They derive from ectomesenchymal cells
originated by migration of neural crest cells during the early craniofacial development. Odontoblasts form the dentine, a
collagen-based mineralized tissue, through secretion of its collagenous and noncollagenous organic matrix components and
by control the mineralization process. A conspicuous cell process arises from the cell body of odontoblasts and penetrates
into the mineralized dentine. After dentinogenesis, odontoblasts deposit new layers of dentine throughout life and might also
form a type of reactionary/reparative dentine in response to dental caries and other external factors may affect teeth.
© 2004 Elsevier Ltd. All rights reserved.
Keywords: Odontoblasts; Dentine; Cell differentiation; Dentinogenesis; Mineralization

Cell facts
• Odontoblasts form the dentine.
• Odontoblasts may deposit new layers of reactionary/reparative dentine.
• Odontoblasts may play a role in dentine sensitivity.

1. Introduction of dentine throughout life. In addition, newly differ-

Odontoblasts are the cells responsible for the for-
mation of dentine, the collagen-based mineralized tis-
sue that forms the bulk of teeth. Odontoblasts derive
from ectomesenchymal cells, exhibit a tall columnar
shape, and establish a continuous single layer with
a clear epithelioid appearance. After dentinogenesis,
they are aligned along the periphery of the dental pulp
thus playing a role in the maintenance of the tooth in-
tegrity owing to their capacity of depositing new layers
∗ Corresponding author. Tel.: +55-11-3091-7308;
fax: +55-11-3091-7402.
E-mail address: vearana@usp.br (V.E. Arana-Chavez).
entiated odontoblast-like cells may also form a layer
of reparative dentine after some tissue injuries. Since
dentine is a tissue analogous to bone, its extracellular
matrix shares many similarities with the bone matrix.
Thus, whereas collagen type I is the major organic
component, the extracellular dentine matrix also con-
tains a variety of noncollagenous proteins. Odonto-
blasts synthesize and secrete all the matrix constituents
and therefore they exhibit well develop synthesis or-
ganelles. The odontoblast layer is separated from the
mineralized dentine by a 10–40 ␮m-thick layer of un-
mineralized matrix, the predentine, which is similar to
the osteoid that separates the osteoblasts/bone lining
cells from the bone mineralized matrix. However, the

1357-2725/$ – see front matter © 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biocel.2004.01.006
1368 V.E. Arana-Chavez, L.F. Massa / The International Journal of Biochemistry & Cell Biology 36 (2004) 1367–1373
odontoblast differs from the osteoblast by possessing
a single cell process that arises from its tall colum-
nar cell body and penetrates into the dentinal tubule.
Thus, the formed dentine is crossed by thousands of
dentinal tubules and canaliculae, which contain the
odontoblast processes and their branches for playing
a role in communication with the dentine matrix; the
cell bodies form the odontoblast layer at the periph-
ery of dental pulp. Although the exact mechanisms on
dentine sensitivity and dental pain are still unknown,
it is believed that odontoblasts play a key role either
by establishing a dentinal/predentinal compartment in
which the dentinal fluid is confined, thus permitting
a hydrodynamic phenomenon; other evidences, how-
ever, consider the possibility of odontoblast might be
a direct sensory receptor or even they may conduct the
nervous impulses to neural pathways.
2. Cell origin
Odontoblasts are derived from embryonic connec-
tive cells that are called ectomesenchymal cells be-
cause of their well-established origin from the neural
crest. Cells from the cranial neural crest early migrate
to populate the first branchial (pharyngeal) arch re-
gions forming the ectomesenchyme that gives arise to
all the soft and hard connective craniofacial tissues.
First, the primitive oral epithelium thickens and prolif-
erates into the underlying ectomesenchyme that under-
goes a conspicuous cellular condensation, which re-
sults in the establishment of dental papilla. A variety of
regulated and reciprocal epithelial-ectomesenchymal
interactions are responsible for the initiation of tooth
formation (Cobourne & Sharpe, 2003). When odonto-
genesis starts, although signaling continues, prolifera-
tion of epithelial cells is a noticeable event occurring
at the bud, cap, and bell stages of odontogenesis. The
differentiation of odontoblasts begins at the tip of the
future cusps only after the tooth germ has reached the
bell stage and the shape of the crown has been estab-
lished.
The ectomesenchymal cells of dental papilla exhibit
a high nucleus to cytoplasm ratio. They possess sev-
eral short processes and contain relatively abundant
polyribosomes but few cisternae of rough endoplasmic
reticulum (RER), Golgi saccules and mitochondria.
Signaling molecules arise from the epithelial cells as
they start their differentiation into ameloblasts, per-
haps modifying the inner basal lamina (bl) in order to
induce the post-mitotic outer cells of the dental papilla
to differentiate into odontoblasts (Ruch, Lesot, &
Begue-Kirn, 1995). Some growth factors, as TGF-1,
BMP 2 and 4, as well as IGF, have been shown to be
expressed by the epithelial cells at this early moment
of development in which the carefully orchestrated
sequence of epithelial-mesenchymal interactions con-
tinues (Thesleff, 2003). It is also possible that some
small amounts of amelogenin molecules that are se-
creted by the differentiated ameloblasts or amelogenin
isoforms or even peptides may participate in that in-
ductive phenomenon (Veis, 2003). The differentiating
odontoblasts start the secretion of dentine before the
initiation of enamel matrix secretion by ameloblasts.
3. Functions: the formation of normal and
reactionary/reparative dentine
Dentine is the mineralized tissue that constitutes the
bulk of the tooth. It is intimately related to the den-
tal pulp with which it shares the same embryological
origin from the dental papilla. As mentioned above,
deposition of the extracellular dentine matrix begins
when the outer cells from dental papilla are induced to
differentiate into odontoblasts. Then, they start their
polarization while elongating and developing synthe-
sis organelles. Differentiating odontoblasts establish a
cylindrical shape and the nucleus locates at the prox-
imal pole of the cell body (Fig. 1). The Golgi ap-
paratus becomes located in the supranuclear region
while numerous reticulum endoplasmic cisternae oc-
cupy the majority of the distal cytoplasm. As polar-
ization and differentiation of odontoblasts start, few
small and short processes arise at the odontoblast sur-
face facing the enamel organ. Differentiating odonto-
blasts then secrete the organic matrix for establishing
the first dentinal layer—the mantle dentine. The earlier
unmineralized matrix is mainly composed by collagen
fibrils, which are deposited into the preexisting ground
substance of the dental papilla. The mantle dentine
collagen fibrils that are conspicuously large and long,
align to form right angles to the basal lamina. As the
differentiating odontoblasts deposit collagen and other
mantle dentine components, numerous small, spheri-
cal, membrane-limited bodies, 50–150 nm in diameter
 V.E. Arana-Chavez, L.F. Massa / The International Journal of Biochemistry & Cell Biology 36 (2004) 1367–1373 1369

Fig. 1. Light micrograph from a tooth germ at the early stage of dentinogenesis. The differentiation of odontoblasts (O) and the onset of
dentine deposition are gradually more advanced from the left to the right side of the micrograph. Odontoblasts exhibit a tall columnar
shape, with their nucleus located at their basal region. Whereas at early stages the forming matrix is unmineralized (asterisk), when it
becomes mineralized (arrows), a layer of newly formed matrix (the predentine, Pd) is always interposed between the odontoblasts and the
mineralizing front. A, differentiating ameloblasts. Hematoxylin-eosin staining (900×).

that are known as matrix vesicles (MV), bud off from
their distal end. It is owing to their small size, the ma-
trix vesicles are only visible under the transmission
electron microscope where they show a variable amor-
phous content (Fig. 2). They establish a close relation
to proteoglycans/glycosaminoglycans that are thought
to bind calcium ions. The first mineral crystals appear
within matrix vesicles, as occurs in immature bone and
in calcifying cartilage (Bonucci, 2002). In the mean-
time, the collagen fibrils, as well as the rest of the
mantle dentine extracellular matrix remain devoid of
mineral.
As differentiating odontoblasts secrete the first lay-
ers of mantle dentine, they reach around 30–40 ␮m in
length, become more closely packed and develop nu-
merous gap and adherents junctions, thus establishing
a cellular layer with a well defined epithelioid appear-
ance. The differentiating odontoblasts begin to move
toward the center of the dental papilla whereas one
of the cell processes gradually becomes accentuated.
This cell process that is left behind into the forming
dentine matrix constitutes the odontoblast process
around which the dentine matrix mineralizes, thus
forming a dentinal tubule. As each odontoblast leaves
behind one single cell process, thousands of denti-
nal tubules cross the dentine from the dentinoenamel
junction to the pulp. The odontoblast process lacks
cell organelles but contains numerous longitudinally
arranged actin filaments and microtubules (Fig. 3).
At stages in which most matrix vesicles are fully
mineralized, odontoblasts appear taller (50–60 ␮m
in length) than at initial stages and focal tight junc-
tions assemble between them. Different from those in
epithelia, the tight junctions between differentiating
odontoblasts play a role that is more related to the
crescent cell polarization and differentiation than to
permeability functions (Arana-Chavez & Katchburian,
1997). Recent studies have shown that the tight
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Fig. 2. Electron micrograph of a region of early dentine mineralization showing numerous matrix vesicles (MV), some of them containing
fine mineral crystals (arrowheads), among the organic mantle dentine matrix rich in collagen fibrils (C). At these stages, the basal lamina
(bl) is partially degraded, while the differentiating ameloblasts (A) present some short projections towards the developing mantle dentine.
The differentiating odontoblasts also possess cell processes (P), which appear sectioned in different directions (26,000×). The inset shows
a high-resolution post-embedding colloidal gold immunocytochemical preparation for dentin matrix protein 1 (DMP 1). The gold particles
are present around a mineralization foci of the mantle dentin in a more advanced stage than that of the large micrograph, in which the
mineral is spreading from matrix vesicles to the surrounding matrix (28,000×).

junctions between differentiating odontoblasts con-
tain more ZO-1, a cytoplasmic junctional protein that
is close related to actin filaments, than occludin and
claudins, two classes of transmembrane proteins that
are related to sealing properties of tight junctions
(João & Arana-Chavez, 2003,2004). Nevertheless,
with the assembly of tight junctions between odonto-
blasts an apical domain is established on their plasma
membrane and they become fully differentiated. Ma-
trix vesicles are no longer released and the secretion
of several noncollagenous matrix proteins begins
(Arana-Chavez & Katchburian, 1998). The differen-
tiated odontoblasts that exhibit a exuberant secreting
apparatus (Fig. 4) continue to deposit a somewhat
different dentine—that is called circumpulpal, which
forms the bulk of the whole dentine. In circumpul-
pal dentine, the collagen fibrils are thinner and they
are more densely packed than those found in mantle
dentine, many of them running around the dentinal
tubules. The noncollagenous dentine matrix proteins
synthesized and secreted by the fully differentiated
odontoblasts include dentine sialophosphoprotein
(DSPP)—a large parental protein that subsequently
suffers cleavage to originate two products, dentine
sialoprotein (DSP) and dentine phosphoprotein (DPP),
dentine matrix proteins 1, 2, and 3 (DMP-1, DMP-2,
and DMP-3, respectively), and small amounts of four
noncollagenous proteins (osteopontin, bone sialopro-
tein, osteonectin, and osteocalcin) that are abundant
in bone (Butler & Ritchie, 1995; He, Dahl, Veis, &
George, 2003; Papagerakis et al., 2002;). Because
mineralization of circumpulpal dentine occurs with-
out the presence of matrix vesicles, it is likely that
noncollagenous matrix proteins play a key role to
establish favorable conditions for the mineral depo-
sition spreading through the fibrilar and interfibrilar
regions of matrix (Fig. 2, inset).
The dentine formed by secretion of collagen and
noncollagenous proteins is called intertubular dentine.
After formation of mantle dentine, each odontoblast
 V.E. Arana-Chavez, L.F. Massa / The International Journal of Biochemistry & Cell Biology 36 (2004) 1367–1373
Fig. 3. Electron micrograph showing the mineralized den-
tine/predentine/odontoblast layer interface. Whereas the mineral-
ized layer of dentine (MD) exhibits higher electron opacity, the
underlying unmineralized predentine (Pd) exhibits numerous col-
lagen fibrils. The density of collagen in predentine gradually de-
creases from the mineralization front, where there is no matrix
vesicles, to the lower regions, in which there are abundant proteo-
glycans and other noncollagenous matrix components. An odon-
toblasts processes (P) that arises from the odontoblast cell body
(O) crosses the entire predentine and penetrates into a tubule of
the mineralized dentine. Some branches (double arrowheads) of
the odontoblast processes are clearly observed in other odontoblast
process (5400×).
secrete additional noncollagenous components from
the end of its cell process. This matrix mineralizes
rapidly between the previously formed intertubular
dentine and the odontoblast process and constitutes the
peritubular dentin. Peritubular dentine is hyperminer-
alized in relation to intertubular dentine and forms the
wall of the dentinal tubule. It is owing to the peritubu-
lar dentine is secreted throughout life of the odonto-
blast, it occurs a gradual obliteration of the dentinal
tubules with aging.
The dentine formed up to the completion of root
development is defined as primary dentine. Because
1371
Fig. 4. Electron micrograph showing the Golgi region of a secretory
odontoblast in which well-developed synthesis organelles such as
cisternae of rough endoplasmic reticulum (RER), Golgi stacks
(G), and secretory granules (g) are abundant. M, mitochondria
(32,500×).
the odontoblasts deposit dentine throughout life, a sec-
ondary dentine is laid down at a much slower rate than
the primary dentine. Both primary and secondary den-
tine posses a similar structure. Formation of secondary
dentine mainly occurs at the roof and floor of the pulp
chamber, thus leading to an increasing reduction of the
pulp volume, as well as of the height of pulp horns.
At these stages the odontoblasts exhibit less developed
synthesis organelles than at stages of primary dentine
formation. It is important to say that the odontoblasts
are post-mitotic cells that remain for tooth life.
Odontoblasts might, however, form a third type
of dentine—the tertiary dentine. There are two sub-
types of tertiary dentine because sometimes the orig-
inal odontoblasts die and then a new generation of
odontoblast-like cells forms a new dentinal layer.
Thus, the tertiary dentine formed by the original
odontoblasts in response to attrition, caries or other
stimuli such as some restorative procedures is called
1372 V.E. Arana-Chavez, L.F. Massa / The International Journal of Biochemistry & Cell Biology 36 (2004) 1367–1373
reactionary dentine. In general, the tubules of re-
actionary dentine continue with those of secondary
dentine, while the thickness of the newly formed
layer is related to the intensity and duration of the
stimulus. The reactionary dentine possesses an or-
ganic matrix as well as a mineral content similar
to those found in primary and secondary dentine.
In other cases, however, some external factors such
as dental caries may irreversibly affect the odon-
toblast layer in a given region. Cavity preparation
during restorative procedures in dentistry may also
affect the odontoblast layer. Whereas shallow cav-
ities produce disruption of the junctions between
odontoblasts, deep cavity preparation has been shown
to cause aspiration of the odontoblast body up into
the dentinal tubule thus disrupting its cytoplasmic
content and resulting in cell death. Although these
situations ultimately promote several pulp patholo-
gies, when they have a reversible behavior, newly
differentiated odontoblast-like cells may arise to
forming another subtype of tertiary dentine that is
called reparative dentine (Smith et al., 1995). It
is assumed that undifferentiated ectomesenchymal
cells present at the pulp, especially at the subodon-
toblastic layer, become the cells that produce the
reparative dentine. Although the exact mechanisms
that induce the differentiation of pulpal cells into
odontoblast-like cells are still unknown, the new cells
arise after a growth factor signaling that triggers a
cascade of events involving cell division, chemotaxis,
cell migration, cell adhesion and cytodifferentiation.
Reparative dentine is in the majority of cases quite
different morphologically from reactionary dentine.
It may contain cellular inclusions, which resemble
the osteocytes from bone. In addition, their extracel-
lular matrix contains some noncollagenous proteins
(osteopontin, for example) that are more typical for
bone than of dentine. For these reasons, sometimes
the reparative dentine shows an osteodentine appear-
ance. It is important to say that differentiation of
pulpal cells into odontoblast-like cells takes place in
absence of enamel organ epithelium and basement
membrane. It is believed that cells from the subodon-
toblastic layer—and perhaps other undifferentiated
pulpal cells—may acquire a latent capability for
this specific differentiation during the morphogenetic
epithelial-ectomesenchymal interactions that took
place at the bell stage of odontogenesis, despite some
components of dentine matrix have also been in-
volved in these mechanisms. Nevertheless, it appears
that absence of inflammation and sufficient oxygen
supply within the pulp may improve the promotion
of odontoblast-like cells cytodifferentiation (Tziafas,
1995).
4. Associated pathologies
Several genetic disorders exhibiting an autosomal
dominant pattern of inheritance may affect the dentine
formation by odontoblasts. They are classified into
two main groups, dentinogenesis imperfecta and den-
tine dysplasia, both with several subtypes. Dentino-
genesis imperfecta type I (DGI-I) is associated with
osteogenesis imperfecta that is caused by mutations
of genes coding for pro-␣ 1(1) chains and pro-␣2(1)
chains of type I collagen, which occur on the long
arm of chromosomes 7 and 17. These mutations re-
sult in helix-destabilizing glycine substitutions, single
exon-skipping mutations, and premature stop codons,
which produce shortened and elongated collagen fib-
rils. In DGI-I, the dentine is hypomineralized and
the overlying enamel is frequently brittle and easily
fractures from it. The dentinogenesis imperfecta type
II (DGI-II) is considered the classical dentinal type;
dentinogenesis imperfecta type III (DGI-III) is also
restricted to the dentine and is the most severe but
it appears to affect restricted populations from the
south of Maryland, USA. Both DGI-II and DGI-III
are not collagen defects but two disorders of den-
tine mineralization; the basic defects appear to be
originated within the human chromosome 4, on the
gene cluster 4q21, from which the main noncollage-
nous dentine matrix proteins DSPP and DMP 1 and
2, as well as the novel protein—matrix extracellu-
lar phosphoglycoprotein/osteoblast-osteocyte factor
45 kDa (MEPE/OF45)—are encoded. Dentine dyspla-
sia, on the other hand, is a rarer disease characterized
by the obliteration of the pulp chamber and by re-
sulting in teeth with short and cone-shaped root(s);
the mineralization in the coronal dentine is normal or
sometimes higher. Some disorders related to the gene
expression of the noncollagenous matrix proteins, es-
pecially DSPP, are likely to play a role in the etiology
of dentine dysplasia (MacDougall, Simmons, Gu, &
Dong, 2002).
 V.E. Arana-Chavez, L.F. Massa / The International Journal of Biochemistry & Cell Biology 36 (2004) 1367–1373
Acknowledgements
The authors would like to thank Sabrina S. Tessi
and Gaspar F. de Lima by their technical assistance as
well as Fapesp and CNPq (Brazil) by their financial
support during the studies on odontoblast biology.
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